TRANSEPITHELIAL TRANSPORT AND ENZYMATIC DETOXIFICATION OF GLUTEN IN MACAQUES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们描述了幼年恒河猴的一种面筋敏感性状况，这种状况在许多方面与人类的面筋敏感症--乳糜泻相似。因此，这种对面筋敏感的动物模型可能有助于研究乳糜泻的发病机制和治疗。在这里，我们进行了两个初步实验，以证明该模型在研究肠道对免疫毒性面筋多肽的通透性和口服酶类候选药物体内谷蛋白的药理解毒方面的潜在用途。研究了年龄匹配的面筋敏感猕猴和对照猕猴的肠道通透性，使用质谱仪检测和定量口服剂量的同位素标记的33聚谷蛋白多肽通过肠道上皮传递到血浆中。一种有治疗前景的口服蛋白酶EP-B2在对面筋敏感的猕猴身上进行了保护效果的评估，方法是每天服用面筋挑战，同时补充或不补充EP-B2。对这只猕猴和年龄匹配的对照组进行了基于ELISA的抗体检测和盲法临床评估，以评估对面筋蛋白的反应。在一只面筋敏感型猕猴的血浆中检测到标记的33肽，无论是在缓解期还是活动期，但在健康对照组的血浆中没有检测到。给予EP-B2，而不是赋形剂，可以防止因饮食面筋挑战而导致的临床复发。出乎意料的是，在EP-B2治疗阶段，抗醇溶蛋白(Ig G和Ig A)和抗组织转谷氨酰胺酶(Ig G)抗体显著增加。对面筋敏感的恒河猴可能是一个有吸引力的资源，用于研究乳糜泻的重要方面，包括增强肠道通透性和口服酶候选药物的药理学。口服EP-B2具有防止面筋摄入的保护作用。有限的数据表明，胃肠道活性谷氨酸酶产生的短面筋多肽的渗透性增强可能会引发抗体反应，但这些抗体并不一定会导致临床疾病。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We characterized a condition of gluten sensitivity in juvenile rhesus macaques that is similar in many respects to the human condition of gluten sensitivity, celiac disease. This animal model of gluten sensitivity may therefore be useful toward studying both the pathogenesis and the treatment of celiac disease. Here, we perform two pilot experiments to demonstrate the potential utility of this model for studying intestinal permeability toward an immunotoxic gluten peptide and pharmacological detoxification of gluten in vivo by an oral enzyme drug candidate. Intestinal permeability was investigated in age-matched gluten-sensitive and control macaques by using mass spectrometry to detect and quantify an orally dosed, isotope labeled 33-mer gluten peptide delivered across the intestinal epithelium to the plasma. The protective effect of a therapeutically promising oral protease, EP-B2, was evaluated in a gluten-sensitive macaque by administering a daily gluten challenge with or without EP-B2 supplementation. ELISA-based antibody assays and blinded clinical evaluations of this macaque and of an age-matched control were conducted to assess responses to gluten. Labeled 33-mer peptide was detected in the plasma of a gluten-sensitive macaque, both in remission and during active disease, but not in the plasma of healthy controls. Administration of EP-B2, but not vehicle, prevented clinical relapse in response to a dietary gluten challenge. Unexpectedly, a marked increase in anti-gliadin (IgG and IgA) and anti-tissue transglutaminase (IgG) antibodies was observed during the EP-B2 treatment phase. Gluten-sensitive rhesus macaques may be an attractive resource for investigating important aspects of celiac disease, including enhanced intestinal permeability and pharmacology of oral enzyme drug candidates. Orally dosed EP-B2 exerts a protective effect against ingested gluten. Limited data suggest that enhanced permeability of short gluten peptides generated by gastrically active glutenases may trigger an elevated antibody response, but that these antibodies are not necessarily causative of clinical illness.